Proteomic studies of B16 lines: involvement of annexin A1 in melanoma dissemination.
To identify proteins involved in melanoma metastasis mechanisms, comparative proteomic studies were undertaken on B16F10 and B16Bl6 melanoma cell lines and their subsequent syngenic primary tumours as pulmonary metastases were present only in the mice bearing a B16Bl6 tumour. 2DE analyses followed by MALDI-TOF identification showed variations of 6 proteins in vitro and 13 proteins in vivo. Differential expressed proteins in tumours were related to energy production and storage. Two differentially expressed proteins which had not been previously associated to melanoma progression, annexin A1 (ANXA1) and creatine kinase B (CKB), were found both in cells and in tumours. To characterize ANXA1 involvement in melanoma B16 dissemination, we reduced ANXA1 protein level by siRNA and observed a significant decrease of B16Bl6 cell invasion through Matrigel coated chambers. We further demonstrated that the presence of several formyl peptide receptors (FPR1, FPRrs1 and 2) revealed by qRT-PCR, played a role in B16 invasion: incubation of B16Bl6 cells with the FPR agonist (fMLP) or antagonist (tBOC) enhanced or decreased Matrigel coated chamber invasion respectively, with a correlation of ANXA1 levels in both treatments. As ANXA1 could bind to FPRs, this should amplify invasion and enhance melanoma dissemination.
Rondepierre
Fabien
F
Imagerie Moléculaire et Thérapie Vectorisée, Rue Montalembert, 63005 Clermont-Ferrand Cedex, France.
Bouchon
Bernadette
B
Papon
Janine
J
Bonnet-Duquennoy
Mathilde
M
Kintossou
Roselyne
R
Moins
Nicole
N
Maublant
Jean
J
Madelmont
Jean C
JC
D'Incan
Michel
M
Degoul
Françoise
F
D000818:Animals; D017305:Annexin A1 / Q000378:metabolism*; D001483:Base Sequence; D045744:Cell Line, Tumor; D003402:Creatine Kinase / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D008546:Melanoma, Experimental / Q000378:metabolism* / Q000473:pathology*; D051379:Mice; D009362:Neoplasm Metastasis; D040901:Proteomics; D044042:Receptors, Formyl Peptide / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014407:Tumor Cells, Cultured
D017305:Annexin A1; D044042:Receptors, Formyl Peptide; D003402:Creatine Kinase